Using Human Pluripotent Stem Cell-Derived Dopaminergic Neurons to Evaluate Candidate Parkinson's Disease Therapeutic Agents in MPP+ and Rotenone Models
Using Human Pluripotent Stem Cell-Derived Dopaminergic Neurons to Evaluate Candidate Parkinson's Disease Therapeutic Agents in MPP+ and Rotenone Models
复制标题
DOI：
10.1177/1087057112474468
复制
发表时间：
2013-06-01
影响因子：
--
通讯作者：
Zeng, Xianmin
中科院分区：
文献类型：
--
作者：
Peng, Jun;Liu, Qiuyue;Zeng, Xianmin
为了开始开发高通量测定系统以评估帕金森病（PD）的潜在小分子疗法，我们使用人多能干细胞衍生的多巴胺能神经元用少量化合物进行了低通量测定。我们首先使用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑鎓测定作为神经保护的读数，在96孔格式的1-甲基-4-苯基吡啶鎓（MPP+）测定中评估了已知在啮齿动物系统中起作用的44种化合物的作用。胶质细胞源性神经营养因子被用作阳性对照，因为其对多巴胺能神经元的神经保护作用已得到充分证实，并对每种药物的两种浓度进行了测试。在筛选的44种化合物中，16种在一种或两种测试剂量下显示出神经保护作用。在MPP+模型中建立了16个阳性子集的剂量-反应曲线。此外，我们验证了这些化合物在鱼藤酮诱导的多巴胺能神经元细胞死亡（另一种已建立的PD模型）中的神经保护作用。我们的人类原发性多巴胺能神经元为基础的测定提供了一个平台，用于快速筛选和/或验证潜在的神经保护剂在PD治疗中使用患者特异性细胞，并显示使用人类细胞进行此类测定的重要性。
To begin to develop a high-throughput assay system to evaluate potential small-molecule therapy for Parkinson's disease (PD), we have performed a low-throughput assay with a small number of compounds using human pluripotent stem cell-derived dopaminergic neurons. We first evaluated the role of 44 compounds known to work in rodent systems in a 1-methyl-4-phenylpyridinium (MPP+) assay in a 96-well format using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay as a readout for neuroprotection. Glial cell-derived neurotrophic factor was used as a positive control because of its well-documented neuroprotective effect on dopaminergic neurons, and two concentrations of each drug were tested. Of 44 compounds screened, 16 showed a neuroprotective effect at one or both dosages tested. A dose-response curve of a subset of the 16 positives was established in the MPP+ model. In addition, we validated neuroprotective effects of these compounds in a rotenone-induced dopaminergic neuronal cell death, another established model for PD. Our human primary dopaminergic neuron-based assays provide a platform for rapid screening and/or validation of potential neuroprotective agents in PD treatment using patient-specific cells and show the importance of using human cells for such assays.